Document Detail


Recombinant λ Bacteriophage Displaying Nanobody towards Third Domain of HER-2 Epitope Inhibits Proliferation of Breast Carcinoma SKBR-3 Cell Line.
MedLine Citation:
PMID:  23224340     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Phage display of many nanobodies via filamentous phage in combination with helper phage has been reported by many scientists. The aim of this study was to produce lambda (λ) bacteriophage displaying high-affinity nanobody against HER-2 expressing breast carcinoma cells. Bacteriophage λ is a temperate phage with inherent biological safety in mammalian cells. Here we report the construction of a recombinant λ phage that efficiently expresses specific nanobody towards third domain of HER-2 target on SKBR-3 and MCF-7 cell lines in vitro. We constructed recombinant λ phage particles containing a mammalian expression cassette, C-Myc tagged, encoding VHH gene of camelid anti HER-2 third domain epitope using λ ZAP-cytomegalic virus (CMV) vector. The SKBR-3, MCF-7 and human endometrial stem cells were treated by the nanobody displayed recombinant λ phage. The cell growth inhibition assay was performed by MTT Cell Viability Assay Kit. After the fourth round of biopanning there was a significant enrichment in the phage specifically binding to the antigen. The ratio of targeted phage increased approximately 1,000-fold in the fifth round. The nanobody expressed by λ ZAP-CMV-VHH phagemid cloned in λ bioparticles significantly inhibited the proliferation of HER-2 positive SKBR-3 and MCF-7 cells. Recombinant bacteriophage λ ZAP-CMV-VHH-cDNA could be used efficiently for construction of nanobodies to mortify HER-2 positive breast carcinoma cells as a nanomedical therapeutic.
Authors:
Alireza Shoae-Hassani; Seyed Abdolreza Mortazavi-Tabatabaei; Shiva Sharif; Shabnam Madadi; Hamidreza Rezaei-Khaligh; Javad Verdi
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-7
Journal Detail:
Title:  Archivum immunologiae et therapiae experimentalis     Volume:  -     ISSN:  1661-4917     ISO Abbreviation:  Arch. Immunol. Ther. Exp. (Warsz.)     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-10     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0114365     Medline TA:  Arch Immunol Ther Exp (Warsz)     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Research Center for Science and Technology in Medicine (RCSTIM), Tehran University of Medical Sciences (TUMS), Tehran, Iran.
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